OP0023 UNBIASED HIERARCHICAL CLUSTERING OF CIRCULATING MOLECULAR PROFILES IN RHEUMATOID ARTHRITIS PATIENTS DEFINES PRECISE CV RISK. EFFECTS OF TNFI, IL6RI, AND JAKINIBS
Authors: L. Muñoz-Barrera, C. Perez-Sanchez, T. Cerdó, R. Ortega Castro, J. Calvo Gutierrez, F. U. Pilar, P. Seguí Azpilcueta, M. C. Ábalos-Aguilera, D. Ruiz, S. Remuzgo Martinez, R. López-Mejías, J. J. Pérez Venegas, M. D. Ruiz Montesinos, C. Dominguez, C. Rodríguez-Escalera, C. M. Romero-Barco, A. Fernandez-Nebro, N. Mena-Vázquez, J. Uceda, C. Santos, N. Barbarroja Puerto, I. Arias de la Rosa, M. A. Aguirre, E. Collantes Estevez, A. Escudero Contreras, M. A. González-Gay, C. Lopez-Pedrera
Journal: Annals of the Rheumatic Diseases (2023)

<h3>Objectives</h3> This study aimed to: 1) Analyze the circulating inflammatory profile of Rheumatoid Arthritis (RA) patients, in order to recognize distinctive clinical phenotypes associated with CV risk; 2) Evaluate modulatory effects of TNF inhibitors (TNFi), JAK-STAT inhibitors (JAKInibs) and IL-6R inhibitors (IL6Ri); 3) Characterize underlying molecular mechanisms involved in endothelial dysfunction. <h3>Methods</h3> Two hundred and eight RA patients and 45 healthy donors (HD) were recruited. Serum inflammatory profile was assessed by analyzing 27 cytokines/chemokines -Luminex assays-, and biomolecules related to NETosis and oxidative stress -by base-plate kits-. Parallel extensive clinical analyses were performed. TNFi, JAKinibs and IL-6Ri effects were evaluated, respectively, in 45, 20 and 17 RA patients after 6 months. Lastly, mechanistic <i>in vitro</i> studies were developed in cultured endothelial cells (ECs) and changes in protein expression were evaluated in cell lysates by proximity extension assay technology, analyzing a panel of 88 proteins related to CV disease. <h3>Results</h3> Unsupervised-clustering identified 3 clusters representing specific molecular profiles. Clinically, even in the presence of similar disease score (DAS28) and positivity for autoantibodies, cluster 1 (C1) identified RA-patients expressing high inflammatory mediators' levels, the highest CV-risk score, and a preponderance of atheroma plaques. Conversely, RA-patients conforming C3 showed the lowest inflammatory profile and the lowest CV-risk score. Lastly, C2 characterized an intermediate phenotype. Comparative analyses with a cohort of 98 RA patients presenting previous CV events, demonstrated that their inflammatory profile mimicked that found in C1, supporting the association of this altered shape with the CV status. <i>In vivo</i>, both biological and targeted-synthetic DMARDs' therapy reduced DAS28-score and re-established normal levels of several altered biomolecules, reflecting a key role in the CV-risk control. <i>In vitro</i>, RA patients' serum pool from cluster 1 promoted in cultured ECs increased expression of several CV-related proteins, further prevented-albeit in an specific way- by the pre-incubation with TNFi (Etanercept), JAKinibs (Baricitinib) and anti-IL6Ri (Tocilizumab). <h3>Conclusion:</h3> 1. The systemic inflammatory profile of RA identified patients' subgroups with enhanced CV-risk, not associated with their disease activity status. 2. Both biological and targeted-synthetic DMARDs re-established normal levels of circulating inflammatory biomolecules, reducing the CV-risk in RA. 3. <i>In vitro</i> studies revealed that RA-serum inflammatory mediators directly induced endothelial damage which might be prevented by effect of both, biological and targeted-synthetic DMARDs' therapy. Thus, the analysis of the RA patients circulating molecular profile might contribute to improve the personalized clinical management of these patients and their CV risk. <h3>Acknowledgements</h3> Supported by ISCIII (PI21/0591, CD21/00187 and RICOR-RD21/0002/0033), and Junta de Andalucía (P20_01367) co-financed by FEDER; Fundacion Andaluza de Reumatología (FAR). <h3>Disclosure of Interests</h3> None Declared.